tbuckley/cabot-search huggingface dataset:
Abstract 2573: A novel regulation of the Wnt/β-catenin pathway involving NLRP12 during colorectal tumorigenesis
Authors: Shahanshah Khan, Lan Peng, Cheryl M. Lewis, Yunpeng Gao, Ram S. Mani, Hasan Zaki
Journal: Cancer Research (2023)

Abstract NLRP12, a member of the NOD-like receptor family, has been characterized as a negative regulator of NF-κB and MAPK pathways in immune cells. We previously demonstrated that Nlrp12-deficient mice are highly susceptible to azoxymethane plus dextran sodium sulfate induced colorectal tumorigenesis in mice. However, the precise mechanism of NLRP12-mediated protection against colorectal cancer (CRC) was poorly understood. RNA-seq analysis revealed that tumors of Nlrp12-deficient mice express higher levels of protooncogenes (cMyc, Ccnd1), Wnt target genes (Ctnnb1, Axin2, Yap1, Tcf3, Lgr5), matrix metalloproteinases, and epithelial to mesenchymal transition markers (Vim, Fn1, Foxc2, Zeb2, Ezh2) compared to wild-type tumor. Interestingly, higher expression of these genes in Nlrp12-deficient tumors was not linked to any inflammatory signaling pathways as there was no difference in the activation of NF-kB, ERK, JNK, AKT, and STAT3 signaling pathways between Nlrp12-/- and wild-type tumors. However, we observed higher activation of β-catenin in Nlrp12-deficient tumors. Using intestinal epithelial cell specific Nlrp12-conditional knockout mice we showed that Nlrp12 deficiency in intestinal epithelial cells leads to increased tumorigenesis, metastasis, and β-catenin activation. To further understand the role of NLRP12 in the regulation of the Wnt/β-catenin, we overexpressed NLRP12 in HEK293T cells and HCT116 cells followed by stimulation with Wnt ligand Wnt3A. NLRP12 overexpressed cells showed significantly reduced activation of β-catenin and expression of Wnt target genes. Consistently, knocking down of NLRP12 in HEK293T and HCT116 cells caused higher activation of the Wnt/β-catenin pathway. Further biochemical analysis revealed that NLRP12-mediated suppression of β-catenin activation is associated with reduced phosphorylation of GSK3β. Interestingly, human and mouse colorectal tumors showed reduced expression of NLRP12 along with increased activation of β-catenin and GSK3β phosphorylation. Altogether, these data suggest that NLRP12 is a potent negative regulator of the Wnt/β-catenin pathway in colorectal tumor cells, and the NLRP12-GSK3β axis could be a novel target for CRC treatment. Citation Format: Shahanshah Khan, Lan Peng, Cheryl M. Lewis, Yunpeng Gao, Ram S. Mani, Hasan Zaki. A novel regulation of the Wnt/β-catenin pathway involving NLRP12 during colorectal tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2573.
Abstract 781: Clinical implications for returning polygenic risk information for skin cancer to ultra-high risk organ transplant recipients
Authors: Mathias Seviiri, Matthew H. Law, Peter Hopkins, Catherine M. Olsen, David C. Whiteman, Stuart MacGregor
Journal: Cancer Research (2023)

Abstract Introduction: Organ transplant recipients (OTRs) have an elevated risk of developing keratinocyte cancer (KC). We have previously shown that a KC polygenic risk score (PRS) effectively triages OTRs for KC risk, with 50% of transplantees with high genetic risk developing KC within 3 years of follow up. However, readiness for clinical application of the KC PRS, its application to triage OTRs in the clinic and the return of the genetic risk information have not yet been assessed. Objectives: 1) To evaluate patients’ readiness to receive genetic risk information for KC risk stratification in a lung transplant clinic. 2) To assess whether returning the PRS results to patients influences skin cancer screening, incidence, and sun protection behavior. Methods: We recruited 100 lung transplantees from the Prince Charles Hospital in Queensland into the ASCOT prospective study. We assessed their attitude, knowledge, and perceptions of genetic testing for KC risk stratification. We also collected genetic data. We developed a KC risk stratification pipeline, and triaged the transplantees. PRS results and KC prevention advice were returned to the high risk group. All transplantees are being followed-up for sun protection and screening behaviors, and skin cancer outcomes. Results: 50% of OTRs were highly likely to accept genetic testing for KC risk stratification. Following transplantation, there was a drastic change in the perception of KC risk and severity, and the majority of transplantees considered it a major health concern. Using the PRS pipeline, 40 patients were triaged into the high risk category for KC, and received their PRS results and health advice, and we are prospectively assessing whether this will influence skin cancer screening rates, skin cancer incidence, and sun protection behavior in this group. Findings will be shared. Conclusion: There is positive patient-readiness to receive genetic risk information for KC risk stratification among transplantees. High-risk patients are currently being followed-up for KC outcomes and prevention service uptake including screening, and sun protection behaviors. Citation Format: Mathias Seviiri, Matthew H. Law, Peter Hopkins, Catherine M. Olsen, David C. Whiteman, Stuart MacGregor. Clinical implications for returning polygenic risk information for skin cancer to ultra-high risk organ transplant recipients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 781.
Abstract 3411: Proof of concept study of next generation drug screening platform for glioma patients
Authors: Nam-Gu Her, Amin El-Heliebi, Gi Ju Lee, San Ha Park, Barbara Prietl, Stephanie S. Ahn, Hong Boon Toh, Thomas R. Pieber, Do-Hyun Nam
Journal: Cancer Research (2023)

Abstract Precision medicine endeavors to match patients to therapies mostly on the basis of the genomic alterations in their tumors. However, it has been shown that ‘genomics only’ approach is often insufficient for best drug selection. Functional precision medicine has been recently emerging as it provides directly translatable information on which drug to choose among available drugs. Traditionally, cancer drugs are tested in cancer cell line models, but cell lines cannot represent individual clinical patients and are too biologically pronounced to be useful for drug screening purposes. Our patient-derived cell (PDC) drug screening system overcomes these points and can generally also correlate with genome changes. AVATAMED and CBmed, together with AimedBio, are collaborating in a Precision Cancer Project, set out next generation drug screening platform to screen Austrian glioma patients for proof-of-concept study. Fresh native tissue from both low- and high-grade glioma are collected. Tumor tissue is used for PDC isolation using mechanical and enzymatic tissue dissociation. To preserve the original tumor similarity, tissue is short term cultured under two weeks, and PDCs are seeded and treated with a panel of clinical- and preclinical drugs followed by viability assessment. We have cultured 65 PDCs from both low- and high-grade glioma. Tissue culture success rate and drug screening success rate was &amp;gt;85% suggesting that specimen logistics and tissue processing are well-established and conducted. PDCs were treated with 79 drugs that are commonly used in clinical setting. In order to evaluate the quality of the results, the screening was conducted at two different sites, AimedBio and CBmed, and the data were compared. Overall, high accuracy and reproducibility were confirmed by low Coefficient of Variation (CV) between replicates and high Z-factor between negative and positive controls. In addition, data generated from both sites showed high correlations. Collectively, our next-generation drug screening platform shows the potential to give the best therapeutic options to glioma patients. To this end, we are moving forward use this platform in clinical study. Citation Format: Nam-Gu Her, Amin El-Heliebi, Gi Ju Lee, San Ha Park, Barbara Prietl, Stephanie S. Ahn, Hong Boon Toh, Thomas R. Pieber, Do-Hyun Nam. Proof of concept study of next generation drug screening platform for glioma patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3411.
Abstract 2486: Dissecting the lineage specific roles of BCL-XL in breast cancer metastasis
Authors: Madison E. Carelock, Mo Jiao, Weizhou Zhang
Journal: Cancer Research (2023)

Abstract B-cell lymphoma X-large (BCL-XL) is a member of the BCL-2 protein family and has been known as an anti-apoptotic protein. As a viable target for cancer therapy, we recently found that proteolysis-targeting chimeras (PROTAC, currently under phase I clinical investigation) against BCL-XL kill cancer cells as well as tumor-induced regulatory T cells (TI-Tregs). Interestingly, BCL-XL has been shown to promote breast cancer metastasis via a function independent of its anti-apoptotic function. We reason that BCL-XL-targeting PROTACs can be potentially used to treat metastatic breast cancer - by direct killing of circulating tumor cells at their vulnerable stages and/or by eliciting anti-cancer immunity via T-reg cell depletion. To our surprise, our pilot experiments using two syngeneic breast cancer models showed opposite results with one model showing reduced metastasis by BCL-XL PROTAC treatment and the other model showing increased metastasis. Genetic deletion of BCL-XL (KO) in those two cell lines reproduces the effect of PROTAC on metastasis, suggesting the on-targeting but contrasting function of BCL-XL in breast cancer metastasis. Our central hypothesis is that BCL-XL may play different roles in breast cancer metastasis depending on the genetics/proteomics of cellular cues from different cancer cells. To confirm the function of BCL-XL in different breast cancer cell lines, we generated BCL-XL KO lines from 4 other syngeneic mouse breast cancers. We confirmed that the majority of breast cancer cells depend on BCL-XL for metastasis, including 4T1, AT-3, E0771.lmb etc; whereas only the Py8119 model exhibits an opposite phenotype. For primary tumor growth, AT-3 and E0771.lmb cells grow bigger tumors in the WT group than in the BCL-XL KO group; and 4T1 and Py8119 cells exhibit similar tumor growth rate between WT and KO cells. For metastatic breast cancer therapy, we treated 4T1-tumor bearing mice with BCL-XL-targeting PROTAC that nearly significantly reduces metastatic spread in the lung without significant change in primary tumor growth. In addition, we found a systematic inhibition of TI-Tregs by PROTACs indicating a lineage-specific function of BCL-XL in breast cancer metastasis. We conclude that BCL-XL is a viable therapeutic target for treating breast cancer metastasis, but caution is required as BCL-XL inhibition may be detrimental for certain cancers. Our future effort will focus on the mechanisms explaining why BCL-XL inhibits metastasis in some cancer types, which can be used as an exclusion criterion for BCL-XL-targeting therapy. Citation Format: Madison E. Carelock, Mo Jiao, Weizhou Zhang. Dissecting the lineage specific roles of BCL-XL in breast cancer metastasis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2486.
Abstract 152: Robust establishment and expansion of human fallopian tube organoids in serum-free culture
Authors: Victor Ho, Maggie Chan, Manreet Chehal, Alice Liang, Allen Eaves, Sharon Louis, John Stingl
Journal: Cancer Research (2023)

Abstract High-grade serous ovarian cancer (HGSOC) is the most prevalent subtype of epithelial ovarian cancer. Despite its name, HGSOC originates in the fallopian tube (FT), but metastasizes quickly in the adnexal region and is the most lethal gynecological cancer worldwide. Organoid culture is emerging as a powerful model for studying normal FT cell biology and ovarian cancer. To standardize primary human FT and HGSOC organoid culture, we have developed GyneCult Fallopian Tube Organoid Medium (FTOM), an optimized serum-free medium and a workflow that supports robust and representative FT organoid culture from freshly isolated or cryopreserved primary human FT cells. FT cultures were initiated by seeding 4000 dissociated single cells directly into 20 µL Corning® Matrigel® domes and overlaying them with FT Organoid Medium. After seeding, cultures were maintained with full-medium changes every 3 - 4 days and were split either in a passage ratio of 1:3, or at 4,000 cells per 20 µL of Matrigel®, as single cells every 8 - 14 days. Cultures were analyzed by immunocytochemistry (ICC) to detect secretory markers keratin 7 (KRT7), oviductal glycoprotein 1 (OVGP1), and PAX8, as well as ciliated cell markers acetylated alpha tubulin (TUBA1A) and FOXJ1. Across all donor samples, approximately 16 ± 8% (mean ± SD) of dissociated EpCAM+ FT cells formed 50 - 300 µm diameter cystic organoids within 14 days (n = 8). Cultures can be maintained for at least 5 passages with 14 - 20 cumulative population doublings, at 3 - 4 doublings per passage. ICC analysis confirmed that organoids contain both polarized KRT7+ OVGP1+ PAX8+ secretory cells and acetylated TUBA1A+ FOXJ1+ ciliated cells, indicating multilineage capacity (n = 5). We also tested the compatibility of GyneCult FTOM for growing HGSOC samples, and we observed that 1 of the 3 tumor samples tested generated organoids with a solid and irregular shaped morphology. These cells could undergo a minimum of 5 cumulative population doublings over 5 passages, at 0.5 - 2 doublings per passage; this performance is in line with recently published HGSOC organoid culture methods. These results demonstrate that GyneCult Fallopian Tube Organoid Medium is a robust medium for initiating and culturing FT epithelium as organoids and is a valuable tool for studying FT biology, with the capacity to support HGSOC organoid culture. Citation Format: Victor Ho, Maggie Chan, Manreet Chehal, Alice Liang, Allen Eaves, Sharon Louis, John Stingl. Robust establishment and expansion of human fallopian tube organoids in serum-free culture [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 152.
Abstract 2682: Targeting phosphoglycerate dehydrogenase (PHGDH) in combination with tamoxifen reduces hormone receptor positive breast cancer growth <i>in vivo</i>
Authors: Brian F. Clem, Yoannis Imbert-Fernandez, Belinda Petri, Carolyn Klinge
Journal: Cancer Research (2023)

Abstract Breast cancer afflicts thousands of individuals every year and is clinically diagnosed in various subtypes, including hormone receptor positive (HR+ BC). Significant strides have been made in treating HR+ BC through the development of endocrine therapies, such as tamoxifen, fulvestrant, and aromatase inhibitors. While these have improved the survival outcomes of these patients, many eventually relapse and, ultimately succumb to metastatic endocrine resistant disease. Subsequent work has identified various cellular pathways contributing to endocrine resistance, including alterations in cellular metabolism. We previously demonstrated that de novo serine synthesis (SSP) is increased in endocrine resistant cell types and genetic disruption of SSP enzymes impacts tamoxifen sensitivity. Further, elevated transcript expression is inversely correlated with survival outcomes in HR+ BC patients treated with tamoxifen. From this, we aimed to assess whether pharmacological targeting of SSP activity in combination with tamoxifen affects the growth of HR+ BC in vivo. We confirmed the presence of ER-alpha in three distinct patient derived xenograft (PDX) models and found varying levels of both phosphoglycerate dehydrogenase (PHGDH) and phosphoserine aminotransferase (PSAT1), which are the first and second enzymatic steps in the SSP pathway, respectively. In our efficacy studies, we initially found differential sensitivity to tamoxifen administration across the PDX models. While treatment with a PHGDH inhibitor (PHGDHi) had no anti-tumor activity against an endocrine-sensitive PDX, it moderately reduced the growth of endocrine resistant PDX tumors. Notably, combinatorial treatment resulted in a substantial decrease in tumor growth in all three models compared to single agents alone. Together, these data suggest that targeting of SSP activity in combination with endocrine therapies may be a promising therapeutic strategy in HR+ BC. Citation Format: Brian F. Clem, Yoannis Imbert-Fernandez, Belinda Petri, Carolyn Klinge. Targeting phosphoglycerate dehydrogenase (PHGDH) in combination with tamoxifen reduces hormone receptor positive breast cancer growth in vivo [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2682.
Abstract 3249: Circulating cytokine associations with clinical outcomes in melanoma patients treated with checkpoint inhibitors
Authors: Jiajia Chen, Giuseppe Tarantino, Mariano Severgnini, Joanna Baginska, Anita Giobbie-Hurder, Michael Manos, Janice D. Russell, Jason L. Weirather, Kathleen Pfaff, Amy Y. Huang, Scott J. Rodig, Srinika Ranashinge, David Liu, F.Stephen Hodi
Journal: Cancer Research (2023)

Abstract Background: aCTLA-4/aPD-1 combination therapy has significantly improved clinical outcomes in patients with metastatic melanoma, with 50%-60% of patients responding to treatment, but predictors of response are poorly characterized. In this study, we set out to evaluate the hypothesis that circulating cytokines and peripheral white blood cell composition may predict response to therapy. Methods: We quantified cytokines and complete blood counts from 89 patients with advanced melanoma treated with combination therapy from three points in time: pre-treatment, one month and approximately three months after starting therapy. We evaluated the association between peripheral blood circulating features, cytokines and clinical labs values, and clinical outcome using predictive modeling, and investigated the prognostic values of these features using survival analysis. Results: Pre-treatment, patients with durable clinical benefit (DCB) had higher IL23, lower CXCL6, and lower IL10 levels. The ratio of neutrophils to lymphocytes (NLR) pre-treatment was not associated with response, but higher NLR one month after starting therapy predicted poorer progression free survival (PFS) and overall survival (OS), primarily driven by an increase in absolute lymphocytes. A multivariate model demonstrated that baseline CXCL6, IL10, IL23 were independent predictors of therapy response, and the combined model has reached an area under the curve (AUC) of 0.8 in prediction of response to combination therapy. Low IL10 and CXCL6 identified an excellent prognosis group, while high CXCL6 and low IL23 characterized a very poor prognosis group of patients. Conclusions: Baseline measurement of cytokines IL23, CXCL6, and IL10 can predict metastatic melanoma patients’ response to ipi/nivo combination treatment. Higher neutrophils-to-lymphocytes ratio approximately one month after therapy started associates with worse survival outcome. Our study presented circulating features from a relatively accessible and less costly avenue, peripheral blood, to be predictive of response to combination immune checkpoint blockade. Citation Format: Jiajia Chen, Giuseppe Tarantino, Mariano Severgnini, Joanna Baginska, Anita Giobbie-Hurder, Michael Manos, Janice D. Russell, Jason L. Weirather, Kathleen Pfaff, Amy Y. Huang, Scott J. Rodig, Srinika Ranashinge, David Liu, F.Stephen Hodi. Circulating cytokine associations with clinical outcomes in melanoma patients treated with checkpoint inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 3249.
Abstract 6381: Quantitative multiplex analysis of soluble and cellular immune checkpoint proteins in response to interferon gamma across multiple murine tumor models
Authors: Wen-Rong Lie, Ryan Bucktrout, Sanjukta Chakraborty, Andrea Orlando, Inna Serganova, Christine Kornmeier, James Hoberg, Roberta Zappasodi
Journal: Cancer Research (2023)

Abstract Immune checkpoint proteins are key regulators of the immune system and drug targets of cancer immunotherapy. Immune checkpoint molecules have been found on the cell surface of various tumor cells and immune cells as immune activating or inhibitory receptors and ligands. Recent studies have shown these immune checkpoint proteins can undergo alternative splicing or proteolytic cleavage leading to soluble isoforms. IFNγ is a pleiotropic and multifunctional cytokine commonly released by cytotoxic immune cells. IFNγ production can be enhanced in response to immunotherapy and further modulates the expression of immune checkpoints in the tumor microenvironment. To better understand the immune checkpoint signatures in tumors, we developed a 28-plex mouse checkpoint protein multiplex immunoassay for quantitative analysis of soluble (secreted) and cellular immune checkpoint proteins. Using this immunoassay, we tested 16 mouse cell lines of tumor (n=13) and normal tissues as control (n=3): 2 melanomas (B16F10, YUMM1.7), 3 mammary carcinoma (4T1, NT5, AT-3), 1 lung carcinoma (LLC), 1 prostate cancer (MycCaP), 1 fibrosarcoma (MCA205), 1 ovarian cancer (ID8), 1 bladder cancer (MB49), 1 glioma (CT-2A), 2 blood cancer (RAW, EL4), 1 dendritic cells (DC2.4), 1 myoblast cells (C2C12), and 1 endothelial cells (bEnd.3). We analyzed changes in immune checkpoint protein expression and secretion in these cell lines cultured in normal conditions or upon exposure to IFNγ to reflect an immune inflamed microenvironment. Cells were cultured in 96-well plate, 100 mm, or 150 mm Petri dishes, and treated with either 10 ng/ml or 100 ng/mL mouse IFNγ. Cell culture supernatants and cell lysates were harvested after 6h, 12h, or 24h incubation with IFNγ and processed for multiplex Luminex bead-based immunoassay. The soluble and cellular checkpoint proteins most substantially modulated by IFNγ treatment in these cell lines included: CD137, BTLA, HVEM, PD-L1. The tumor-specific signature of cellular and soluble checkpoint proteins and its response to IFNγ may have implications in the tumor microenvironment and in the outcome to immunotherapy. Citation Format: Wen-Rong Lie, Ryan Bucktrout, Sanjukta Chakraborty, Andrea Orlando, Inna Serganova, Christine Kornmeier, James Hoberg, Roberta Zappasodi. Quantitative multiplex analysis of soluble and cellular immune checkpoint proteins in response to interferon gamma across multiple murine tumor models [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6381.